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目的观察环磷酸腺苷(c AMP)类似物二丁酰环磷腺苷(db-c AMP)对大鼠矽肺纤维化的保护作用。方法雄性Wistar大鼠随机分为3组(n=10):对照16周组皮下注射生理盐水10 mg/(kg·d),矽肺模型16周组皮下注射生理盐水10 mg/(kg·d),染尘至16周,db-c AMP预防治疗组皮下注射db-c AMP 10 mg/(kg·d),染尘至16周。采用Masson三色染色观察肺组织病理形态的改变,Western blot检测α-平滑肌肌动蛋白(α-SMA)、Ⅰ型胶原(CollagenⅠ)、纤连蛋白(Fn)、Gαs、Gαi2/3和c AMP的含量,利用免疫荧光化学染色检测矽肺组织α-SMA/Gαi3的共表达。结果 Masson染色显示,结节及间质纤维化区域出现蓝紫色胶原沉积。免疫荧光结果显示,纤维化病变区域α-SMA/Gαi3共表达增强,矽肺模型16周组与对照16周组比较,Gαs和c AMP的表达明显下降,Gαi2/3及CollagenⅠ、α-SMA、Fn的蛋白表达明显升高。与矽肺模型16周组相比,db-c AMP预防治疗组肺组织胶原沉积减轻,α-SMA/Gαi3共表达下降,Gαs和c AMP的蛋白表达明显升高,Gαi2/3及α-SMA、CollagenⅠ、Fn的表达显著降低。结论 db-c AMP对大鼠矽肺纤维化具有保护作用,这与其调节Gαs/Gαi信号促进c AMP的生成,抑制肌成纤维细胞转化及细胞外基质的生成有关。
Objective To observe the protective effect of cAMP analogue db-c AMP on silicotic fibrosis in rats. Methods Male Wistar rats were randomly divided into 3 groups (n = 10): the control group was subcutaneously injected with saline 10 mg / (kg · d) for 16 weeks, and the saline group was injected subcutaneously with saline 10 mg / (kg · d) , Dust to 16 weeks, db-c AMP prophylaxis treatment group, subcutaneous injection of db-c AMP 10 mg / (kg · d), Dust to 16 weeks. Masson trichrome staining was used to observe the pathological changes of lung tissue. Western blot was used to detect the expression of α-SMA, Collagen Ⅰ, Fn, Gαs, Gαi2 / 3 and c AMP The expression of α-SMA / Gαi3 in silicosis tissue was detected by immunofluorescence staining. Results Masson staining showed blue-violet collagen deposition in nodules and interstitial fibrosis areas. The results of immunofluorescence showed that the expression of α-SMA / Gαi3 in fibrosis lesions was enhanced. Compared with the 16-week control group, the expression of Gαs and cAMP in the silicosis model group decreased significantly. The expressions of Gαi2 / 3 and CollagenⅠ, α-SMA, Fn The protein expression was significantly increased. Compared with the 16-week group, the protein expression of Gαs and c-AMP in the db-cAMP group was decreased, the expression of α-SMA / Gαi3 was decreased, the expression of Gαi2 / 3 and α- Collagen Ⅰ, Fn expression was significantly reduced. Conclusion db-c AMP has a protective effect on silicotic fibrosis in rats, which is related to the regulation of Gαs / Gαi signal to promote the production of cAMP, inhibit the transformation of myofibroblasts and the formation of extracellular matrix.